【目的】观察支气管上皮细胞(BEAS-2B细胞)和嗜中性粒细胞(Neutrophils细胞)接触共培养体系中基质金属蛋白酶组织抑制剂-2(tissue inhibitors of metalloproteinases-2,TIMP-2)合成,探讨不同剂量葛根素对共培养体系TIMP-2的抑制作用。【方法】应用蛋白芯片筛查不同培养体系炎性细胞因子表达;应用ELISA方法定量测量TIMP-2浓度;应用Real-time PCR方法检测TIMP-2基因表达。【结果】BEAS-2B细胞和Neutrophils细胞联合培养上清液中TIMP-2浓度升高,基因表达水平明显上调(P<0.05)。经葛根素干预可显著抑制两种细胞联合培养TIMP-2浓度及基因表达(P<0.05)。【结论】Neutrophils细胞与BEAS-2B细胞联合培养可显著增加细胞培养上清液中炎性细胞因子TIMP-2浓度,上调BEAS-2B细胞中TIMP-2基因表达;葛根素可下调TIMP-2浓度及表达。
Objective:In this study,we aimed to investigate the expressions of adhesion molecules on human bronchial epithelial cells and neutrophils in co-culture system,assess the effects of puerarin on suppressing these adhesion molecules expressions,and explore the roles of two crucial signal-transduction elements p38mitogen-activated protein kinase(p38 MAPK)and nuclear factor kappa B(NF-k B)in modulating adhesion molecules expressions.Methods:Neutrophils and BEAS-2B cells(one human bronchial epithelial cell line)were co-cultured,and adhesion molecules expressions on cell surface were detected using flow cytometry.The mRNA levels of adhesion molecules were assessed by real-time quantitative polymerase chain reaction(real-time qPCR).Phosphorylated p38 MAPK and inhibitor k B were analyzed by Western blot.Results:In co-culture system,adhesion molecules expressions on BEAS-2B cells and neutrophils were enhanced significantly(P<0.05).Correspondingly,the mRNA levels of adhesion molecules were also increased greatly.Moreover,the pretreatment of peurarin obviously suppressed adhesion molecules expressions on cell surface.Furthermore,phosphorylated p38 MAPK and inhibitor k B in BEAS-2B cells and neutrophils were elevated in co-culture system,but decreased significantly after upon the treatment of peurarin(P<0.05).Conclusions:Coculture boosted the interactions between human bronchial epithelial cells and neutrophils mimicking airway inflammation,whereas peurarin decreased the expression of adhesion molecules on cell surface by suppressing the activities of d38 MAPK and NF-κB pathways,and exhibiting its anti-inflammation activity.
